Chapter title |
An Organotypic 3D Assay for Primary Human Mammary Epithelial Cells that Recapitulates Branching Morphogenesis
|
---|---|
Chapter number | 9 |
Book title |
3D Cell Culture
|
Published in |
Methods in molecular biology, June 2017
|
DOI | 10.1007/978-1-4939-7021-6_9 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7019-3, 978-1-4939-7021-6
|
Authors |
Jelena R. Linnemann, Lisa K. Meixner, Haruko Miura, Christina H. Scheel M.D., Christina H. Scheel |
Editors |
Zuzana Koledova |
Abstract |
We have developed a three-dimensional organotypic culture system for primary human mammary epithelial cells (HMECs) in which the cells are cultured in free floating collagen type I gels. In this assay, luminal cells predominantly form multicellular spheres, while basal/myoepithelial cells form complex branched structures resembling terminal ductal lobular units (TDLUs), the functional units of the human mammary gland in situ. The TDLU-like organoids can be cultured for at least 3 weeks and can then be passaged multiple times. Subsequently, collagen gels can be stained with carmine or by immunofluorescence to allow for the analysis of morphology, protein expression and polarization, and to facilitate quantification of structures. In addition, structures can be isolated for gene expression analysis. In summary, this technique is suitable for studying branching morphogenesis, regeneration, and differentiation of HMECs as well as their dependence on the physical environment. |
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Geographical breakdown
Country | Count | As % |
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Unknown | 42 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
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Student > Ph. D. Student | 6 | 14% |
Researcher | 6 | 14% |
Student > Bachelor | 5 | 12% |
Student > Doctoral Student | 4 | 10% |
Student > Master | 2 | 5% |
Other | 3 | 7% |
Unknown | 16 | 38% |
Readers by discipline | Count | As % |
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Medicine and Dentistry | 3 | 7% |
Immunology and Microbiology | 2 | 5% |
Pharmacology, Toxicology and Pharmaceutical Science | 2 | 5% |
Other | 2 | 5% |
Unknown | 19 | 45% |